The primary objective of this project is to further demonstrate the feasibility of developing recombinant human milk fat globule epidermal growth factor-factor 8 (rhMFG-E8) as a novel and effective adjuvant therapy for the resuscitation of patients with hemorrhagic shock. Hemorrhagic shock is a major cause of mortality worldwide. In the United States, traumatic injury is the main cause of death in individuals younger than 44 years of age, and hemorrhagic shock accounts for one third of trauma-related deaths. Even when treated with the standard of care and in a hospital setting, 10% of patients with hemorrhagic shock die within hours. Therefore, there is an unmet and critical need for a novel and effective adjuvant therapy capable of improving the success rate of the existing volume and vasoactive agents for hemorrhage resuscitation. MFG-E8 is a protein that promotes the clearance of apoptotic/dying cells and inhibits the production of pro-inflammatory cytokines. In preliminary studies, we treated hemorrhaged animals with rhMFG-E8 and observed significant decreases in circulating pro-inflammatory cytokines, neutrophil infiltration to the lungs, and apoptosis. Treatment with rhMFG-E8 also nearly doubled the survival rate from 43% to 83%. Therefore, we hypothesize that rhMFG-E8 can be further developed as a new and effective adjuvant therapy for hemorrhagic shock. To advance the drug development, we will produce a new non His-tagged rhMFG-E8 and analyze its biological activity, homogeneity, and folding status. We will then determine the dose-dependent effects of rhMFG-E8 on reducing organ injury, hemodynamic instability, pro-inflammatory cytokines, and histological damage caused by hemorrhagic shock. Its therapeutic window to improve survival after hemorrhagic shock will be investigated. Finally, we will determine its safety pharmacology and pharmacokinetic profile in healthy and hemorrhaged animals. Our future steps (SBIR Phase II and beyond) will include completing preclinical studies, such as ADME studies and efficacy studies in a non-rodent species, as well as comparing with the standard of care. We will then file an investigative new drug (IND) application with the FDA to initiate clinical trials. Our ultimate goal is to obtain commercial utilization of rhMFG-E8 as a safe and effective adjuvant drug for the resuscitation of patients with hemorrhagic shock.